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The lower convective layer (LCL) at Atlantis II brine pool of the Red sea represents one of the exceptional, unique ecosystems. Thioredoxin is a multi-functional antioxidant redox protein that has a crucial role in various vital cellular processes. In the current study, a functional metagenomics approach was used to isolate and characterize thioredoxin from the LCL of Atlantis II Deep brine pool (Trx-ATII). From the metagenomic DNA of the LCL, the thioredoxin gene was directly retrieved and sequenced. Sequence analysis showed that the gene belonged to thioredoxin-like superfamily with classical Trx motif (-CXXC-). Phylogenetic analysis revealed that Trx-ATII was closely related to Trx of Prochlorococcus marinus with a maximum identity of 86%. Successfully, Trx-ATII was cloned and expressed in E. https://www.selleckchem.com/products/gsk963.html coli, where the purified protein had M.wt of 16 kDa. Characterization studies revealed that Trx-ATII protein is halophilic; can tolerate up to 2.5 M NaCl and thermostable, where 90% of its activity was retained at 60 °C. Trx-ATII can reduce both DTNB and insulin disulfide- containing substrates. In conclusion, a unique thioredoxin protein was isolated from a harsh environment that can maintain its activity under extreme conditions of salinity and temperature as a promising redox protein for biotechnological applications. Prenylated flavonoids are good drug candidates due to multiple biological activities and health benefits. Prenyltransferase is an important enzyme involved in the biosynthesis of prenylated flavonoids. In this work, a flavonoid prenyltransferase (AhPT1) from Artocarpus heterophyllus showed an unexpectedly metal ion-induced specificity to flavonoid substrates. AhPT1 could catalyse 6-C-prenylation of genistein when Mg2+ serving as cofactor. Its catalytic activity to 6-hydroxyflavone was undetectable. However, when Mn2+ was used instead of Mg2+, 5-C-prenylated 6-hydroxyflavone was generated with a high conversion rate. Mn2+ altered the regiospecificity of AhPT1 and turned it into a 5-C-prenyltransferase. 2'-Hydroxyl could improve the conversion rate of 6-hydroxyflavone, whereas 3'- or 4'-hydroxyl impaired the catalysis efficiency of AhPT1. NQIFDADID174 and DLTDVEGD305 were active motifs involved in substrate binding and catalysis. Asn166, Asp170, Asp174, Asp298, Asp301 and Asp305 in the active center were critical to the prenylation reaction. A composite film composed of whey protein isolate (WPI) and psyllium seed gum (PSG) was investigated. Its physicochemical, mechanical and structural properties were determined at different ratios of WPI/PSG (10, 31, 11, 13, 01). WPI/PSG composite films had higher water contact angle and water vapor permeability, as well as lower oxygen permeability and light transmittance as compared with single WPI or PSG films. With the increase in PSG concentration, higher film brightness and whiteness index, and smaller total color difference were observed. The tensile strength and elastic modulus of the composite film at WPI/PSG ratio of 11 was the highest. Elongation at break of composite films was higher than that of the single films. WPI/PSG composite films were more effective than single films in reducing the surface cracks and degree of cracks. XRD revealed a typical semi-crystalline amorphous structure of the composite films. With the increase of the PSG content, higher diffraction peak strength and crystallinity of the films were observed. The results indicated that the properties of the WPI/PSG composite film were superior to that of PSG or WPI film alone. The composite film at WPI/PSG ratio of 11 resulted in the highest comprehensive physicochemical and mechanical performance. The phycocyanin was purified by Sephadex- G-100 and RP-HPLC and protein content was found to be 52.82% and the high purity fraction was collected and RP-HPLC analysis of fractionated phycocyanin, the α-subunit and β-subunit were detected in 4.9 and 11.1(mAU). The frequency of peak 1456.26 cm-1 has showed the CH2 bending vibration and the protein amide II band was detected at 1539.20 cm-1 (CO stretching) and 2358.94 cm-1. In 1H NMR analysis, 14 chemical shifts (δ) were observed and signals confirmed namely alkyl halide, alkene, aldehyde proton and carboxylic acid. The in vivo anticancer effect was assessed by MTT assay against HepG-2 cell lines and in vivo antidiabetic effect was carried out through α-amylase and β-glucosidase enzyme inhibition methods. The promising anticancer effect 68% was noticed at the concentration of 500 μg/ml and lower anticancer effect was noticed at the concentration of 100 μg/ml against Hep-G2 cell lines. The α-amylase and β-glucosidase enzyme inhibition of phycocyanin showed dose dependent and maximum inhibition effect at 250 μg/ml. Phycocyanin anti-inflammatory effect such as inhibition of albumin denaturation, antiproteinase, hypotonicity-induced haemolysis and anti-lipoxygenase activities have been recorded maximum level at 500 μg/ml. Phycocyanin have complex structure and high molecular weight with more biomedical applications for drug development. Biopolymers of gellan gum (G), 2-hydroxyethyl cellulose (HEC), and lignin (L)-based binary and ternary sustainable composites were prepared for food packaging and biomedical application. The composite films were flexible and transparent or translucent with slight brown in color. The incorporation of lignin considerably improved the thermal and mechanical and hydrophobic properties of the composite films. The addition of 10 wt% of lignin to the composites increased the tensile strength by 54.3% and 59.2% respectively. The prepared lignin-based composite films showed high ultraviolet (UV) protection, with almost 100% protection against UVB (280-320 nm) and 90% against UVA (320-400 nm). The surface hydrophobicity of the composite films increased with the addition of lignin. The binary and ternary composites containing 1, 5, and 10 wt% lignin exhibited excellent radical scavenging activities. The gellan gum/HEC/lignin based composite films achieved the best biocompatibility. The obtained composites showed efficient antioxidant and non-cytotoxic activities, although there was no remarkable antimicrobial activity.
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